Bisnorallo-7-cholenaldehydes



Unitsd SWSP Q ,i i

2,895,970 BISNORALLO-l-CHOLENALDEHYDES Gerald ll). Laubach, Jackson Heights, N.Y., assignor to Chas. Pfizer & Co., Inc., Brooklyn; N.Y., a corporation of Delaware No Drawing. Application September 24, 1954 Serial No. 458,268

1 Claim. (Cl. 260-397.5)

This invention relates to novel stei oid type and their preparation. More particularly, it; concerns new and useful intermediates in the 'syiithe'sis of steroid hormones like cortisone. This present application a continuation-in-part of parent application Serial, No. 2 33,4 8 9, fi led on lune 25, 1951 and abandoned.

Several of the steroid-type contitiientsof the adrenal cortex have been shown to be of great use in the therapy of certain diseases like rheumatoid arthritis. The volume of cortisone and other such compounds which can be eprepared commercially has been severely limited by the short supply of certain naturally occurring materials,-sucli as the bile acids, which have been the necessary starting reactants in the usual manufacturing process. It has recently been found possible to utilize other steroid-type raw materials which are more available, e.g. ergosterol.

However, one problem encountered in the course of in synthesizing cortison this new kind of synthesis is cleavage of the long side.

. stituents.

It has been found that steroid compounds possessing a nuclear double bond at the 7,8-carbon position and a double bond at the 22,23-carbon position may be treated in such a manner as to cleave selectively the C17 side chain and leave a three carbon fragment at this position. The three carbon fragment may then be further degraded to a fragment of the same two carbon chain length as occurs in the especially valuable cortisone and related adrenal cortex hormones. I

The process of this invention, whereby 'splittingfaiid gle gradation of the C17 side chain of a 7 ,8unsaturat ed' steroid is accomplished selectively at the 22-carbon atom, broadly comprises treating the steroid with ozone and reductively cleaving the ozonide thus formed to obtain the corresponding aldehyde. The yield of product from this reaction is good despite the instability of some aldehydes. There may be a minor amount of further oxida tion of the side chain or slight cleavage of the steroid nucleus, but not to a degree which materially interferes with the efiiciency of the process. The products recovered are steroids having. a three-carbon aldehydic substituent at the C17 position, i.e. compounds of the gen eral structure:

ong-on-ono CH3 H30 2,895,970 ....?e*srts4.-lslx 5 2 wherein Ris a inember 'of thegroup hydrogen; allgyl'aiid acyl. A varietyof alkyl groups, e.g; me thyl etfll rid benzyl are useful. The safn i e' is true for he cyl gr p, which has been ae'etyr, p dpionyl, butyryl ljngojl. These changes in. the gr p at the" 3 positing" havno eifecfon the overall real on. Inthisformula in all others in this application, h' dotted line" Hiatfth 5 position is used the conventional sense; lie. 'toshow the 2110 configuration;

To degrade this side chain still further then converted into an acylated eiiol or eiiol ester type of structure; i.e. of the formula:

ga eamete wherein R is chosen the group hydrogen, alkyl and acyl and R is acyl. This agyl group been any of a wide variety, changes here making no difference in the overall reaction. The useful groups include, for example, acetyl, propionyl, butyryl and benzoy'l. This inturn is oxidized to the corresponding ketone. Steroids naiing the particularly desired two-carbon side chain at the HI wherein R is chosen from the group hydrogenfa'lliyl afid acyl. These compounds are especially valuable for conversion to biologically active cortisone, Compound F and like cortical hormones. i l

Rather than degrading. the three carbon fragment at.

the 17-positions of these steroid comppunds to a two carbon fragment, compounds having a three carbon carboxylic acid group may be prepared:

oxidation here.

k enyl side chain at the 17-position may be oxidized (eg.

with ozone) directly to a carboxylic acid of the type described above, there is a tendency during this reaction to attack the nuclear double bond. I prefer to form the aldehyde and then oxidize the aldehyde by means of a mild oxidizing agent to the acid. Ozone may be used for the latter reaction but the quantity must be controlled. The acids may be'converted to the corresponding esters by conventional methods. Although the steroid acid having a free hydroxyl group at the 3-position may be prepared, it may be preferred to retain an alkyl or acyl group attached to the oxygen at this position to prevent The reactions of this invention may be summarized by the following diagram in which S-dihydrbergosteryl acetate (I) is used as an'exaniple of suitable starting material. (I) is ozonized and reduced with cleavage to obtain (II); this is enolizedand esterified, e.g. treated with acetic anhydride, to product (HI); and (III) is oxidized in any desired way to recover (IV). (II) is oxidized to the acid (V).

H CHr-CCH=CH H o H CH. 1 Hc--c-oH,

(I) 5-dlhydroergoatery1aeetate (IV) sp-acetoxyallnJpregnen-w-one These compounds are all very valuable, particularly as intermediates for the synthesis of steroid hormones such as cortisone. For example, the Compound IV above,

'3/3-acetoxyallo-7-pregnen-20-one, is selectively dehydrogenated to introduce a 9(11) double bond, according to -the method of my copending application, Serial No. 45 8,-

269, filed simultaneously herewith, and now abandoned. This product is then converted by the method of Stork et al., J.A.C.S., vol. 73, p. 3546 (1951), to 3,8-ol-allopregnane-11, 20-dione-3-acetate, which in turn is converted to cortisone by the method of Chemerda et al.,

' I.A.C.S., vol. 73, p. 4052 (1951).

Compounds that are particularly useful as starting ma- I 1 l terials for these reactions are the 7,8-mono-unsaturated steroids having the general structure:

of these are a-spinasterol, its ethers and esters, and chondrillasterol (7,8; 22,23 poriferastadienel), its ethers and (II) 3B-aeetoxybisnorallo-7-cholenaldehyde (I11) Enol acetate of II esters, (The latter substance is a C epimer of aspinasterol.) Typical formulas are given below:

One may readily use estersof such sterols, e.g. the acetate as previously noted, the propionate or the benzoate. Alternatively the methyl, ethyl or other ethers may be employed. 'It' rageaerany advisable to protect the 3- hydroxy group of the sterol by formation of an ester or ether, in smarts revent oxidation at 'this 'point of the nucleus. It is a "peculiar fact that the nucleardouble bon at them; position is not at all arrested by the oxidatioii. A 5,6-un'satur'a'tiori, for instance, will generally iseeessit'aterhe addition of bromine or a similar 'pr'otec- 'tix'feagnt Ito the r'ac'tionniass, but zne'noes not alter the riuelear struc ore of a '7,8 m6no-un 'saturated steroid. 'Ihisis the'm'ainreasoh why the latter type of compound is" preferred for iise in this inventioii.

selective e slight 'te e be somewhat improved thereby, it is also {generally hetter to 'ihcludein reactioniiiiixture tertiary organic b lilieipyridinej, qhiholine, or triethylamine. The reiig p n may be at veryilow temperatures, such as -5320; but to obtain a jreasgonable rate r reac ion it isjpreefire cl"to per at lat ab'outO" Higher temperasites may be u 'd, ,butthereis a tendency for'decomposition of thereac'taht's and product, p l

Ozone may beprodu'ced'by a t ehpa n be isolated fromgeactiorr mixtures in wh ch more en used: sidej-reactionsmay o'c'dur rely oxidizin the C 2,, Hanna bands with mm a -7.-..- Y .4 acid in these 'solvents, and since {the product-yield seems sult in deciea'sedyie a. On 'the'oth'er'han'd, nan aippfciably lower proportion than 1.5 moles of ozone per mole of starting material is used, then some steroid will be left unreacted.,. Using a standard laboratory ozonizer which produces aboiifofie'iriillimole of ozone per'liter of gasi 's j l 'xx i sii r l e .r or "one liter. per r'riinut'e fwith r c t applied,', the. average laboratory run 39% .ys n P tminute. nd. sta ti w t rre s of "'h'the" ours ,The, oz d oxygen 'is generally passed o s intered page; the te race/argue can; .mi twleiy h is e a lsby' esx fii iih ster.sz ni t tipn-fiee-bss j e s t th Me i Preferabl s qv redb QQPs n -s the ts s ,rnixture i n vacuum uiider nitrogen. Generally the teniper a ture is hept below about C.to preventdecompositi9 ,...s ,tl 91 .g met-aeria 'con ia'er' ue ity- .Ih o id thus. bta n t st nre for tleavege mi reduction; totem thetles d e de... sc n ft one specific embodiment,ofithe,invention theozonide a I J H 75 active "compounds, such as' c'or'ti'sone. Rather thanusing is'dissolved in a mixture of'acetic acid and an inert sols vefit', sueh "as ether, and the's'olution is cooled to about 0: C. While a stream of nitrogen is passed through. the mixture, fzi'n c dust is added to effect the decomposition. rjthis 'reaeton has been completed, ejg. when pero'xide tests are negative, further solvent is add d and the solution is washed with dilute sodium bicarbonate and then with Water. The water is removed and the dried solvent solution concentrated toyield a partially crystalline aldehydic product. This 'may be purified "if desired, for example bxtdturafing w th a low r, al QllQL Pr a y methanol, at. room rs htlx h p e u Any nnrea edstar am teri l s not h sb .sli e ltsfl-lm alcoho ex ra tofithe aldehyd mayths be sq pt tdm emative m e prqdu tma b f h P r ed by fl rtiingthesodium hisulfite adduct of the aldehyde. I solatimer thisma eriaL-asa o id .a lr v s q t h aldeh de ive armd stfif u tshishp i xt lit-i urally possible to use other methods for conyertingp the ozonide to ,tl 1 corresponding aldehyde, instead of this zinc. du -acid..re qtio r. tanse Q P Y Q r fi sqn methods max h pl ed, i JJ P S ibl'? l tiQllSQ fQI such reactions will be obvious to those skilled .in the,, rt'. M, Q H, s 4 A. Esters. of, the enolizedform of the aldehydes thus produc dare advant g ous x pr ed y, t ti th eldehydewith asu t bl as l t n omma -V. Fo inst ss, referringtgthe reaction series hereinbefore described, the .35ea etoxyb nb al azlz o al hyde (II); marks heated w th cet c nhx rids i in .f s dlsgd um 0 acetatetonm ue exqellentyiel o t e rre pond n enol acetate (III). The enol ester is finally oxidized to the.corresp in twixc rbpn chze sketc e -1 V) in heafqr a dfi. ydteer oster l a at ies. :Su sta ,ti lxanxpxid in a neral y u e u f r. such lreaction may be employed. However, ozone is ;particglarly su tah nf c nver n the 0 4 to, h k t ne i Q d ;y d-....Wh n th a e 9 t syn s s ha l ee reac d, t e e isnQ ava b .a. w ar on cha n-a the. lZ-rcsiflq o the StQ QidYD QIlS- Si ce h sii th same type 9 2.9 7. .subs t eut a fo n in t olo cal y act e Q ti0n .,C mp91 E a t elike, i i apparen that these productQketones serve as excellent intermediates is! h prcp at qn of. su c t ca n tal repared by a, conventionallreaction from the ketones eakest-assesinter d ate 0 The oxidation of the steroid aldehydes to thep'correspe din a idm rbe a ri d ou w th. the use of var'ious tox d ln r snts gene hr u e u o such a r c n-fi h heen fpund that chromic; acid in strong sulfuric acid ution orinjasolution containing, inaddition to sulfuric acid, water-miscible, ,organic solvents which. are stable under th'e'conditionsjofrthe reaction, is particularly ,useful.1,, ,Solve n tssuchas acetic acid or acetonej'are quite useful for this purpose. Furthermore, the'reaction ma ;be frun in the presence. ofa .Water inirriiscible "solvent, such as ether, in which thefaldhydeis dissolved. In this .case,,th e reactionoccursin a two-phase system and strong agitationvisadvisable. For instance, by. meansof this reagent. (chromic acid)", 3fl-acetoxyb-isnorallo r-7-cholenalldehydeIH) has beenconverted to thecorresponding acid (Y). It'is advisable to run' the reaction at a low temfperature, preferably not. over about 5 C., for a short time, at the most several hours. Ingeneral, the reaction may be followed by observing the change in colorwhich occurs during the oxidation. Prolonging the time or raising the temperature may tend to induce side reactions. This acid, and compoundswiththesame nuclear structure ands deph n txaryin in. h na of th group substituted in the 3 -posi'tion (othereSters, ethers, etc), have a definite advantage in that they may be carried through various reactions involving the steroid nucleus d' p ay considerable stability. After such'reactions ornpleted, the '3 carbon side chain may then be deaea to g psu'ai f2 carbon side chain of "biologically the acid, an ester, such as the methyl ester, maybe utilized in carrying out such further reactions. v

The following examples are given by way of illustration and are not to be considered as the sole embodiments of this invention, protection of which is only to be limited by the specific wording of the appended claims.

EXAMPLE I 3fl-acet0xybisnorall0-7-cholenaldehyde An ice cold solution of S-dihydroergosteryl acetate (0.05 mole, melting point 176-182 C.) in 1250 ml. of chloroform and 25 ml. of pyridine was ozonized for 85 minutes (0.085 mole of ozone circulated at the rate of 0.001 mole/liter/ minute at 1 liter/minute). When ozonization was completed, nitrogen was'blown through the cold solution, which was then concentrated under vacuum in a stream of nitrogen at a temperature not exceeding 3040 C.

The oily residue was dissolved in 125 ml. of acetic acid and 250 ml. of ether. Zinc dust (20 grams) was added while the solution was held at C. and agitated by a swirling motion. After minutes of mixing, 1000 ml. of ether were added and the suspension was filtered. The filtrate was extracted with water and then saturated bicarbonate solution, and finally washed with water until neutral. The ether layer was dried over anhydrous sodium sulfate and concentrated at reduced pressure. A tan magma resulted, which on trituration with three 50-ml. portions of methanol yielded 3.91 grams (18%) of unreacted starting material of excellent purity, melting point 164177 C.

The methanolic filtrate was poured as a thin stream into 400 ml. of 30% sodium bisulfite solution to afford an immediate curdy precipitate. After 30 minutes the solid was filtered 0E, washed with 50 ml. of ice water, broken up under 50 ml. of ether, refiltered and washed with ether. When dried overnight in a desiccator, this bisulfite adduct was treated under nitrogen with 1800 ml. of ether and 720 ml. of 10% sodium carbonate solution. The ether layer was separated after standing four hours and the remaining bisulfite compound further treated with 25 grams of sodium carbonate in water and 1000 ml. of ether until all of the solid had disappeared. The combined ether layers, when water washed and dried,

gave 71.2 grams (47% yield) of the crystalline aldehyde,

melting point 125-130 C.

A portion of this aldehyde (0.100 gram) was converted to the semicarbazone by treatment with 0.200 gram each of semicarbazide hydrochloride and sodium acetate in a refluxing methanol-water mixture. An immediate precipitate of 0.086 gram (75%) of the semicarbazone was obtained, melting point 220224 C. One recrystallization gave an analytical sample, melting at 228.8 C. with decomposition. V

Analysis.Calcd. for C H O N C, 69.90; H, 9.15; N, 9.78. Found: C, 69.93; H, 8.83; N, 9.71.

In a manner analogous to the above, this reaction was carried out using a variety of substituents at the 3 position instead of the acetoxy group described above. These changes had no effect on the overall reaction. The used groups included, for example, esters of propionic, butyric and benzoic acids, and the methyl, ethyl and benzoyl ethers. The reaction also went when the 3 position contained the free hydroxyl group.

EXAMPLE II Enol acetate 01'' 3fl-acetoxybisnorallo-7-cholenaldehyde A mixture of 1.86 grams (0.005 mole) of the aldehyde prepared in Example I and 1.0 gram of fused sodium acetate in 50 ml. of acetic anhydride was heated at reflux under nitrogen for 12 hours. After cooling overnight, the crystallized solid was separated by filtraa methanol-water mixture.

tion, washed with water and dried. The product weighed 1.23 grams and melted at 152158 C. Additional solid was recovered by concentration of the mother liquor and dilution with methanol. This weighed 0.15 gram (melting point 146l55 C.) The combined yield was 71%. Recrystallization from ethanol afforded an analytically pure sample of the enol ester as large platelets, melting point 156159 C.

Analysis.Calcd. fOI' Cad-I350: C, H, 9.24. Found: C, 75.23; H, 9.04.

By analogous methods, using acylating agents other than acetic anhydride as described above, other enol esters were prepared from the aldehyde. These included, for example, the propionate, butyrate and benzoate.

EXAMPLE HI 3p-acetoxyallo-7-pregnen-20-one A solution of 1.24 grams (0.003 mole) of the enol acetate from Example II in 35 ml. of ice cold chlorw form was ozonized for 3 /2 minutes (0.0035 mole of ozone, rate 0.001 mole/liter/minute at 1 liter/minute). Nitrogen was blown through the solution, which was then concentrated in vacuo under nitrogen. The residue was dissolved in 20 ml. of ether and 10 ml. of acetic acid and treated with 1.0 gram of zinc dust under nitrogen for 30 minutes. The reaction mixture was diluted with ml. of ether, filtered, and extracted with water, saturated sodium carbonate solution, then with water to neutrality. The solution was dried over anhydrous sodium sulfate and concentrated to produce a crystalline, colorless solid, which on recrystallization from methanol gave 0.760 gram (71% yield) of the desired ketone, melting point 127-145 C.

A highly purified sample was obtained from 1:4 and 2:3 benzene-petroleum ether eluates by chromatography of the ketone over alumina, followed by recrystallization from methanol. This product had the melting point 164166 C. and the optical rotation [0:1 +51 (0., 0.95 in chloroform). The infrared spectrum showed a strong ketonic carbonyl band at 5.85 mg.

Analysis.-Calcd. for C H O C, 77.06; H, 9.56. Found: C, 77.39; H, 9.54.

A semicarbazone was prepared by treating 0.100 gram of the non-chromatographed ketone with 0.200 gram each of semicarbazide hydrochloride and sodium acetate in The yield of crude product, melting at 237 C. with decomposition, was 0.100 gram (86%). Recrystallization from a chloroform-toluene mixture afforded an analytical sample, melting at 268.8 C. with decomposition. I

Analysis.Calcd. for C24H37O3N3: C, 69.36; H, 8.98; N, 10.11. Found: C, 69.26; H, 8.66; N, 10.03.

EXAMPLE IV 3 8-acetoxybisnorallo-7-cholenic acid A solution of 0.373 gram (0.001 mole) of 3flt-acetoxy bisnorallo-7-cholenaldehyde in 10 ml. of ether and 5 ml. of acetone was treated with a solution of 0.0005 mole (0.150 gram) of sodium dichromate dihydrate in 20% sulfuric acid. The mixture was stirred at 0 for'two hours, at which time complete utilization of the oxidant had occurred. The green acid solution was extracted with ether and the combined solvent layers washed with water. Addition of 5 ml. of 5% sodium hydroxide solution afforded a voluminous precipitate of insoluble so dium salt. The basic layer was extracted three times with IO-milliliter portions of ether, then acidified under 10 ml. of ether with 2 N sulfuric acid. The ether layer was washed with water, shaken with sodium sulfate, and finally concentrated under vacuum to afford a 50% yield of nearly pure acid, melting point -l85 C.

What is claimed is: A steroid compound having the structure 2,895,970 r 1 10 wherein R is chosen from the group consisting of hydrogen and hydrocarbon acyl containing up to 7 carbon atoms and R is hydrocarbon acyl containing up to 7 carbon atoms.

References Cited in the file of this patent UNITED STATES PATENTS Heyl et a1 Dec. 23, 1952 Reinhold Feb. 5, 1957 OTHER REFERENCES Bergman et a1.: J. Org. Chem. 13, pages 10-20 (1948). 

